FIG 2.

Localization of proteases in OMVs by proteinase K susceptibility assay. OMVs from C6709 and CHA6.8 ΔprtV were incubated at 37°C for 30 min with or without 1 μg/ml proteinase K alone or with 1% SDS or 1 mM PMSF. Lane 1, control reaction; lane 2, sensitivity to extracellular protease; lane 3, inhibition of extracellular proteases; lane 4, protease sensitivity to luminal proteins; lane 5, inhibition of protease activity by PMSF (control). OMVs were incubated under the above-described conditions, precipitated with acetone, dried, electrophoresed, and immunoblotted against HAP (A) or VesC (B) antiserum.